促血管生成素载体构建及在癌细胞中的表达

Study on construction and expression of eukaryotic plasmid with human angiopoietin-2 cDNA in human cervical hela cells

[目的 ] 构建人促血管生成素 - 2 (Ang - 2 )基因的重组pcDNA3真核表达载体 ,为进一步研究Ang - 2在肿瘤血管生长上的作用奠定基础 .[方法 ] 从人胎盘组织中提取Ang - 2总RNA ,经过RT -PCR扩增、TA克隆、酶切鉴定、DNA序列分析后 ,将纯化的Ang - 2基因克隆到pcDNA3真核表达载体上 ,转染Hela细胞 ,应用RT -PCR方法鉴定细胞表达的mRNA .[结果 ] RT -PCR得到的产物经DNA测序 ,与Genebank中人Ang - 2cDNA序列一致 ;构建人Ang - 2真核表达载体 ,转染人Hela细胞 ,细胞表达Ang - 2目的基因mRNA .[结论 ] 成功构建人Ang - 2基因真核表达载体pcDNA3-Ang - 2 .

Objective] Constructing recombinant eukaryotic expression vector coding for human Angiopoietin-2 in order to investigate the role of Ang-2 in carcinoma angiogenesis . [Methods] Extracting mRNA from human placenta;Identification the gene through RT-PCR,TA clone,restrict enzymes and DNA sequence analysis ;connecting Ang-2 to pcDNA3 and transfecting human hela cell;RT-PCR to identify mRNA expression. [Results] The gene sequence is identical with that reported in genebank.RT-PCR result indicated that recombinant plasmid expressed in hela cell.[Conclusion] Sucessfully constructing Ang-2 eukaryotic express vectors.